JP2670742B2 - α-amylase inhibitor - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION The present invention is applied to guava (scientific name: Psi
(Dium Gugjava Limn; a tropical evergreen small tree belonging to the family Fusariumaceae)) and having a substance having an enzyme inhibitory activity, which is more specifically α
The present invention relates to an extract having α-amylase activity derived from guava leaves having amylase inhibitor activity.

[0002]

BACKGROUND OF THE INVENTION Due to changes in lifestyle, especially eating habits, the symptoms or signs of adult diseases such as hypertension, diabetes, cerebral infarction, heart disease, etc. have spread to the younger generation, causing serious problems. Has been taken up as. In many cases, since such adult diseases are closely related to obesity, improvement of dietary content and elimination or prevention of obesity by exercise are regarded as important as the first prevention thereof. The most common dietary measure is restriction of calorie intake, but this involves changes in the quantity and quality of meals, which can be painful and stressful for many people.
Making it difficult to continue dietary restrictions. Therefore, if it is possible to prevent or delay the ingested food from being metabolized in the living body and being converted into energy, it is possible to obtain an effect of reducing the amount of meal while having a feeling of being full. In particular, substances which inhibit certain starch hydrolysis enzyme is an energy source α- amylase activity is effective in diet, such as wheat and barley [Shiraishi Wuchang, "Chemistry and Biological" 27 49
1-492 (1989)] and laurel (JP-A-4-273).
No. 89) is disclosed as an anti-obesity diet food. However, it is more effective,
There is a strong demand for the development of safe α-amylase inhibitors.

[0003]

Means for Solving the Problems The present inventors have conducted intensive studies with the aim of obtaining a substance having an effective and safe α-amylase inhibitory activity. It was found that the extract thus obtained has a strong α-amylase inhibitor activity, and the present invention has been completed. That is, according to the present invention, the guava leaf is 1.0
The present invention provides a guava leaf extract having an α-amylase inhibitory activity, which is obtained by heating to 100 to 130 ° C. and extracting under −2.0 atm. In order to obtain the extract of the present invention, it is preferable to use water or a hydrophilic solvent or a mixture thereof as an extraction solvent. For the purpose of the present invention, the guava leaf extract having the α-amylase inhibitory activity of the present invention is also simply referred to as an α-amylase inhibitor in the specification.

[0004] In order to prepare an α-amylase inhibitor from guava leaves, conditions are selected under which the extraction efficiency of various components such as polyphenols and saponins contained in guava leaves is maximized. As the hydrophilic solvent, a solvent commonly used in the art, such as ethanol and acetone, can be used. These can be used alone or with one another or in combination with water. A particularly preferred solvent is water. Extraction is optional as long as the α-amylase inhibitor activity of the guava leaf extract is not impaired. Extraction conditions will vary depending on the solvent selected, but usually from normal pressure to under pressure,
That is, heating or heating is performed in the range of about 1 atm to about 2.0 atm. It is preferably carried out under a pressure of 1.2 to 1.7 at or near the boiling point of the solvent. In the case of water, the extraction temperature is about 100-1
30 degreeC, Preferably it is 110-120 degreeC. Under such conditions, extraction is completed in tens of minutes to several hours, generally in tens of minutes, specifically for 30 to 40 minutes. Guava leaves may be raw or dried, but they are all
Cut into a size of about 1 mm to several cm and an average of 5 mm square.
This is put into a pressure type extraction pot and extracted in a usual manner.

After the completion of the extraction, the extract is separated to remove impurities, and then concentrated in an ascending can vacuum concentrator to remove the extraction solvent. Since the obtained guava leaf extract contains the active ingredient contained in the guava leaf at a high concentration, it can be directly added to a liquid or gel food or processed into a pellet or the like. Alternatively, it may be purified by chromatography or the like. The guava leaf extract obtained by the method of the present invention is completely sterilized by various bacteria, especially heat-resistant spores, which may be present in guava leaves as a raw material, has extremely high safety, and has excellent properties as a food additive.

The guava leaf extract obtained as described above is dried by a freeze-dry (high vacuum freeze dryer) or a spray dryer and pulverized to be added to foods, or pellets which are convenient to carry and easy to eat. It is convenient to form granules, tablets or capsules. The guava leaf extract obtained by the method of the present invention exhibits about 3 to 8 times the activity of the existing amylase inhibitor, as shown in the test examples described later, and is useful for inhibiting starch hydrolysis by α-amylase. I found out. Therefore, the present invention provides a method for producing an α-amylase inhibitor, which comprises extracting guava leaves with water, a hydrophilic solvent, or a mixture thereof.

Further, the present invention provides a diet food containing an α-amylase inhibitor derived from guava leaves. The diet food of the present invention can be in any form. For example, in order to easily consume the α-amylase inhibitor as it is, it is formed into a pellet, tablet, capsule or the like together with a suitable base. Alternatively, in order to make it easier to eat, it may be formed into a confectionery shape such as cookies, jellies and snacks. Furthermore, Gymnema sylvestre extract, which is a carbohydrate absorption inhibitor [Hirata et al, Yonago Medical Journal 43
392-396 (1992)] or a peptide that inhibits lipid absorption [Kyoichi Kagawa, Monthly Food Chemical 1
990 (2) 1] and the like.

[0008]

The present invention will be described below with reference to examples. Example 1 About 125 kg of dried guava leaf is crushed to an average of 5 mm square and put in a pressure type extraction pot (capacity 1500 L). Add 600 L of water and heat. The internal temperature of the extraction pot is 120 ° C and the pressure is 1.
Extraction is continued for 30 minutes after reaching 3-1.5 atm. The heating is stopped, the filter cloth is passed through the kettle, and the filtrate is passed through a filter press to remove impurities. Next, the separated extract is subjected to a rising can concentrator to remove water, and 90 L of concentrated guava extract (Brix 25) is obtained. This extract is dried by freeze-drying (high vacuum freeze dryer) or spray dryer to be powdered to obtain about 22 kg of a light brown powdered guava leaf extract.

Example 2 Approximately 125 kg of dried guava leaves were crushed to an average of 5 mm square and put in a pressure type extraction pot (capacity 1500 L). Add 600 L of water and heat to reflux for 30 minutes. The heating is stopped and the product is concentrated and lyophilized in the same manner as in Example 1 above to obtain 19 kg of a light brown powder of guava leaf extract.

Example 3 Approximately 125 kg of dried guava leaf was crushed to an average of 5 mm square and placed in a pressure type extraction pot (capacity 1500 L). Add 600 L of water-ethanol (1: 1) mixture and extract at 60 ° C. for 30 minutes. The heating is stopped, and the product is concentrated and freeze-dried in the same manner as in the above example to obtain about 21 kg of a light brown powdered guava leaf extract.

The extracts prepared in the above Examples were tested for α-amylase inhibitor activity by the following method. Test Example 1 As a substrate solution, a solution obtained by dissolving 0.5% of a soluble starch (manufactured by Matsutani Chemical Industry Co., Ltd.) in 50 mM acetate buffer (pH 6.5) was used.
-Amylase was used. 50 μl of enzyme solution in 0.25 ml of substrate solution, 1 to 10 mg of guava leaf extract powder or laurel extract extract (Nippon Milling Co., Ltd.) obtained in the above example
In 100 ml of water, 50 μl containing 20 mM calcium chloride and 100 mM sodium chloride
50 μl of M acetate buffer (pH 6.5) and 100 μl of 50 mM acetate buffer (pH 6.5) were placed in a test tube and reacted in a water bath at 37 ° C. for 15 minutes. After the reaction is completed, 1.
0.1% containing 7 mM potassium iodide and 0.17 mM iodine.
5 ml of 0017N hydrochloric acid aqueous solution was added, and the absorbance was measured at 700 nm. For comparison, the same components as above except that the enzyme inhibitor solution was not added were put into a test tube and reacted, and used as a control. The OD ₇₀₀ in the control experiment was defined as 100% amylase activity, and the concentration of the test substance that inhibited it by 50% (IC ₅₀ , μg / ml) was determined and compared. Table 1 shows the results.

[0012]

[Table 1] As is clear from Table 1, the guava leaf extract of the present invention has a higher α-amylase inhibitor activity as compared with the conventional extract. In particular, since the extract obtained by high-pressure extraction with water has higher activity, it can be seen that the high-pressure extraction method is particularly suitable in the present invention.

Example 4 Dietary food 1 Guava leaf extract powder obtained in Example 3 3.334 kg Vitamin B ₁ 0.2 kg Vitamin B ₂ 0.2 kg Vitamin B ₆ 0.2 kg Pine flow (base) (Matsutani) Chemistry) 5.766 kg Calcium tertiary phosphate (base) 0.3 kg The above components were put into a compounding machine, stirred for 10 minutes, taken out, and then put on a direct compression tableting machine to have a diameter of 6 mm, a length of 4 mm, and a weight of 150 mg. Make pellets of. This is coated with a thin film of shellac on a shellac coating machine to obtain a product. Each pellet contains 50 mg of guava leaf extract powder. The pellets may be granulated by a granulator.

Example 5 Dietary food 2 Guava leaf extract extract powder obtained in Example 2.0 2.0 kg Gymnema sylvestre extract extract powder 2.0 kg Peptide powder (Hankyu Kyoeisha) 2.0 kg Vitamin B ₁ 0.2 kg Vitamin B ₂ 0.2 kg Vitamin B ₆ 0.2 kg Pine flow (base) 3.1 kg Tricalcium phosphate (base) 0.3 kg The above components were treated in the same manner as in Example 2 to obtain a diameter of 6 mm and a length of 4
mm and weight 150 mg. Guava leaf extract powder 30 mg, Gymnema sylvestre extract extract 30 mg, peptide powder 30 mg per 1 pellet,
Vitamin _B 1 3 mg, containing vitamin _B 2 3 mg, and vitamin _B 6 3 mg. The pellets can also be granulated.

Example 6 Diet food 3 300 g of shortening, 20 g of milk and 50 g of sugar are mixed well with a whisk. Add 60g of eggs little by little and mix well. Separately, 300 g of flour, 1 g of baking powder, and 10.5 g of the guava leaf extract powder obtained in Example 1 are mixed well, and added to the above mixture, followed by further mixing. After leaving it in the refrigerator for 30 minutes, it is divided into about 70 and molded into an appropriate mold.
Bake in the oven for 18-25 minutes. Eating one cookie means eating 150 mg of guava leaf extract.

[0016]

The guava leaf extract of the present invention is a safe and excellent α-amylase inhibitor, and can be used for preventing obesity or adult diseases in addition to diet foods and the like.

Claims (5)

(57) [Claims] 1. Guava leaves under 1.0 to 2.0 atmospheres,
A guava leaf extract having α-amylase inhibitory activity obtained by heating and extracting at 100 to 130 ° C. 2. The guava leaf extract according to claim 1, wherein the pressure at the time of extraction is 1.2 to 1.7 atm. 3. Guava leaves are treated with water or a hydrophilic solvent, or a mixture thereof under 1.0 to 2.0 atm.
Α- characterized by extraction at 00 to 130 ° C
A method for producing a guava leaf extract having amylase inhibitory activity. 4. The method according to claim 3, wherein the pressure during the extraction is 1.2 to 1.7 atm. 5. A diet food containing a guava leaf extract having the α-amylase inhibitor according to claim 1.